The clinical impact of sample storage at -20°C on renin reference intervals and aldosterone-renin ratio calculations.

Cryoactivation was known to occur in whole blood and plasma samples when kept between +4 and -5°C leading to falsely high renin concentrations. In 2022 it has been clearly shown that cryoactivation can also occur in samples stored at -20°C.Based on these new findings here we discuss how this can influence the clinical diagnosis of patients. First, we show that storage of renin plasma samples can affect the renin measurements and thereby the aldosterone-to-renin ratio (ARR) calculation, which might explain the high intra-individual variability in ARR also recently demonstrated. Second, we discuss the existing studies on the establishment of renin reference intervals and noted the lack of attention given to this recently revealed preanalytical conditions. Our literature review of the reference intervals for renin suggest that cryoactivation might have influence the published data.

Case report: Skin protective barrier cream interference in benzethonium chloride method for urine protein measurement in a 6-month-old girl.

This case report describes the positive interference of the commonly used skin protective barrier cream used together with urine collection bags on the benzethonium chloride method for urine protein measurements in a 6-month-old female baby, leading to falsely elevated results. The interference was identified by both artificially mixing urine samples with this cream and comparing the results obtained using the benzethonium chloride method with those obtained using the pyrogallol red method.

Efficacy and Safety of Ibrutinib Therapy in Patients with Chronic Lymphocytic Leukemia: Retrospective Analysis of Real-Life Data

Objective: This study aimed to retrospectively evaluate the efficacy, safety, and survival outcome of single-agent ibrutinib therapy in chronic lymphocytic leukemia patients. Materials and Methods: A total of 136 patients (mean age ± standard deviation: 64.6±10.3 years, 66.9% males) who had received at least one dose of ibrutinib were included in this retrospective multicenter, noninterventional hospital-registry study conducted at 33 centers across Turkey. Data on patient demographics, baseline characteristics, laboratory findings, and leukemia-cell cytogenetics were retrieved. Treatment response, survival outcome including overall survival (OS) and progression-free survival (PFS), and safety data were analyzed. Results: Overall, 36.7% of patients were categorized as Eastern Cooperative Oncology Group (ECOG) class 2-3, while 44.9% were in Rai stage 4. Fluorescence in situ hybridization revealed the presence of del(17p) in 39.8% of the patients. Patients received a median of 2.0 (range: 0-7) lines of pre-ibrutinib therapy. Median duration of therapy was 8.8 months (range: 0.4-58.0 months). The 1-year PFS and OS rates were 82.2% and 84.6%, respectively, while median PFS time was 30.0 (standard error, 95% confidence interval: 5.1, 20.0-40.0) months and median OS time was 37.9 (3.2, 31.5-44.2) months. Treatment response (complete or partial response), PFS time, and OS time were better with 0-2 lines versus 3-7 lines of prior therapy (p<0.001, p=0.001, and p<0.001, respectively), with ECOG class 0-1 versus class 2-3 (p=0.006, p=0.011, and p=0.001, respectively), and with Rai stage 0-2 versus 3-4 (p=0.002, p=0.001, and p=0.002, respectively). No significant difference was noted in treatment response rates or survival outcome with respect to the presence of comorbidity, bulky disease, or del(17p). While 176 adverse events (AEs) were reported in 74 (54.4%) patients, 46 of those 176 AEs were grade 3-4, including pneumonia (n=12), neutropenia (n=11), anemia (n=5), thrombocytopenia (n=5), and fever (n=5). Conclusion: This real-life analysis confirms the favorable efficacy and safety profile of long-term ibrutinib treatment while emphasizing the potential adverse impacts of poorer ECOG performance status, heavy treatment prior to ibrutinib, and advanced Rai stage on patient compliance, treatment response, and survival outcomes.

Erythrocyte zinc level in patients with atopic dermatitis and its relation to SCORAD index.

INTRODUCTION: Atopic dermatitis (AD) is a chronic, pruritic inflammatory disease, characterized by a relapsing-remitting course. The pathogenesis of atopic dermatitis is not completely understood, although the disorder appears to result from the complex interaction between immune abnormalities, genetic and environmental factors. Trace elements are essential for normal functioning of the immune system. AIM: To determine zinc levels in serum and erythrocytes of patients with AD using an atomic absorption spectrometric technique and to investigate the relationship between those levels and disease activity. MATERIAL AND METHODS: Sixty-seven patients and 49 controls were enrolled into the study. The disease severity of AD patients was determined according to the Scoring Atopic Dermatitis (SCORAD) index. We measured zinc levels in serum and erythrocytes by the atomic absorption spectrophotometric technique. RESULTS: Erythrocyte zinc levels were significantly lower in AD patients than in the control group (p < 0.001), whereas serum zinc levels did not differ between the groups (p = 0.148). In the AD patient group there was a negative correlation between the SCORAD score and erythrocyte zinc levels (r = -0.791; p < 0.001). CONCLUSIONS: The negative relationship between disease severity and erythrocyte zinc levels might suggest an immunopathological link between AD progression and intracellular zinc metabolism.

The effect of serum, intestinal and peritoneal visfatin levels on early diagnosis of acute mesenteric ischemia.

BACKGROUND: Acute mesenteric ischemia (AMI) is a rapidly progressive disease where early diagnosis is life-saving. As a new cytokine, levels of thevisfatin might be affected during the ischema and reperfusion. In our study, we obtained changes of visfatin levels in the serum, peritoneal and intestinal lavage samples in rats, to investigate the effectiveness of these changes in the early diagnosis of AMI. METHODS: In group 1 (Sham group) the intestine was exteriorated after the laparotomy was performed and allowed to stand for 3 hours without ischemia. In group 2 (acute mesenteric ischemia-reperfusion group) the mesenteric artery was ligated and, mesenteric blood flow was restored after 60-minute ischemia. To compare with intestinal injury, in group 3 (acute pancreatitis group) the ductus pancreaticus was ligated, and the abdomen was closed for 3 days in expectation of the formation of pancreatitis. In all of the groups, the intestinal lavage, peritoneal lavage and blood samples were analyzed to evaluate the levels of visfatin, TNF-alpha, IL-6 and IL-8. Samples were taken before the procedure in all groups; additionally 60 minutes after ischemia and 120 minutes after reperfusion in group 2; and after the development of the pancreatitis in group 3. RESULTS: Serum, intestinal and peritoneal lavage visfatin levels were found to be increased in group 2 and group 3 (P<0.05). In group 2, while serum TNF-alpha levels were increased in both ischemia and reperfusion; in intestinal lavage sample the increase was only in the ischemic phase (P<0.05). In group 2, IL-8 levels were significantly increased after ischemia in serum (P=0.03) and after reperfusion in intestinal lavage (P=0.004) samples. CONCLUSIONS: Serum, intestinal and peritoneal visfatin levels were increased not only in the case of mesenteric ischemia, but also in acute pancreatitis. In these two clinical pathologies, the visfatin levels of the intestinal and peritoneal cavitiesmay increase parallel to the serum visfatin levels.

Preservative solution for skeletal muscle biopsy samples.

CONTEXT: Muscle biopsy samples must be frozen with liquid nitrogen immediately after excision and maintained at -80°C until analysis. Because of this requirement for tissue processing, patients with neuromuscular diseases often have to travel to centers with on-site muscle pathology laboratories for muscle biopsy sample excision to ensure that samples are properly preserved. AIM: Here, we developed a preservative solution and examined its protectiveness on striated muscle tissues for a minimum of the length of time that would be required to reach a specific muscle pathology laboratory. MATERIALS AND METHODS: A preservative solution called Kurt-Ozcan (KO) solution was prepared. Eight healthy Sprague-Dawley rats were sacrificed; striated muscle tissue samples were collected and divided into six different groups. Muscle tissue samples were separated into groups for morphological, enzyme histochemical, molecular, and biochemical analysis. STATISTICAL METHOD USED: Chi-square and Kruskal Wallis tests. RESULTS: Samples kept in the KO and University of Wisconsin (UW) solutions exhibited very good morphological scores at 3, 6, and 18 hours, but artificial changes were observed at 24 hours. Similar findings were observed for the evaluated enzyme activities. There were no differences between the control group and the samples kept in the KO or UW solution at 3, 6, and 18 hours for morphological, enzyme histochemical, and biochemical features. The messenger ribonucleic acid (mRNA) of ß-actin gene was protected up to 6 hours in the KO and UW solutions. CONCLUSION: The KO solution protects the morphological, enzyme histochemical, and biochemical features of striated muscle tissue of healthy rats for 18 hours and preserves the mRNA for 6 hours.

Methylenetetrahydrofolate reductase (MTHFR) 677C>T gene polymorphism as a possible factor for reducing clinical severity of psoriasis.

Methylenetetrahydrofolate reductase (MTHFR) is a crucial enzyme in homocysteine/methionine metabolism. It catalysis the formation of 5-methyltetrahydrofolate (5-methyl-THF), which is the methyl donor for synthesis of methionine from homocysteine (Hcy). Decreases in folate consumption due to MTHFR polymorphism may affect production rate of keratinocytes of which had faster reproduction rates with a continuous DNA turnover and this may affect the clinical picture of psoriasis. This study aimed to investigate correlation of C677T polymorphisms in the MTHFR gene with severity of psoriasis and to evaluate the status of plasma Hcy, folate and vitamin B12 levels in patient with chronic plaque psoriasis. The study included 60 patients with chronic plaque psoriasis. The C677T polymorphisms were genotyped using PCR (Qiagen). Psoriasis Area and Severity Index (PASI) score below 7 was defined as mild, between 7 and 12 as moderate, and above 12 as severe disease. There was a significant difference between the severity of disease classification (p<0.05) with respect to the C677T polymorphism in the MTHFR gene. Severe involvement (PASI score >12) was observed in 38.46% of wild type (CC), but only 12.50% of homozygote (TT) and 7.69% of heterozygote (CT) patients. Significant differences between gene polymorphism and Hcy levels were noted in TT and CT genotypes respectively (p=0.025 and p=0.040). Plasma Hcy, folate and vitamin B12 levels were not correlated with the PASI score. Our data indicate a possible correlation of MTHFR polymorphism with severity of psoriasis.

Mean platelet volume in children with attention deficit hyperactivity disorder.

The mean platelet volume (MPV), the accurate measure of platelet size, is considered a marker and determinant of platelet function. MPV can be a potentially useful prognostic biomarker in patients with cardiovascular disease. After reviewing literature, we hypothesized that attention deficit hyperactivity disorder (ADHD) in childhood may be a risk factor for coronary heart disease (CHD) in adulthood. The aim of this study was investigation of MPV and platelet count (PLT) in children with ADHD and healthy subjects. The MPV and the PLT were measured in 70 children with ADHD (aged 6-16 years), and compared with 41 healthy controls. The MPV was found to be significantly increased in ADHD group compared to control group (p=.006). There was no significant difference in the PLT between groups (p>.05). To our knowledge, this was the first study of investigating the levels of MPV and PLT in children with ADHD. Although significance and cause of increased MPV level in ADHD remain unclear in present study, further studies are warranted to investigate relationships among MPV, ADHD in childhood and CHD in adulthood.

Folate and homocysteine metabolisms and their roles in the biochemical basis of neuropsychiatry.

The term 'one-carbon metabolism' is commonly used to describe 3 separate metabolic processes: folate metabolism, the homocysteine remethylation cycle, and the transsulfuration pathway. Folate metabolism concerns the biochemical reactions in which endogenous and exogenous one-carbon units are transferred to tetrahydrofolates. The remethylation cycle is used for the synthesis of methionine from homocysteine with one-carbon units that come from folate. This methionine is thenfused for the synthesis of S-adenosyl methionine, which is a general donor of methyl groups for many biochemical reactions in the human body. In the transsulfuration pathway, some amino acids and polypeptides, such as cystathionine, cysteine, and glutathione, are synthesized from homocysteine. The kinetics of the enzymes in this pathway are regulated by the substrates of the remethylation cycle. The methylation process has been thought to have an important role in the biochemical basis of neuropsychiatry. An elevated homocysteine level is the most important marker of folate and vitamin B12 deficiencies, and also the most reliable biochemical sign of functional insufficiency. Some neurological and neuropsychiatric diseases, such as psychosis, Alzheimer's disease, and autism, have been found to be related to disorders of one-carbon metabolism. This review aims to summarize both one-carbon metabolism and its relationships with neuropsychiatric disorders.

Comparison of the effects of first and second generation silicone hydrogel contact lens wear on tear film osmolarity.

AIM: To compare the effects of first and second generation silicone hydrogel (SiH) contact lens wear on tear film osmolarity. METHODS: The healthy subjects who have never used contact lenses before were enrolled in the study. Tear film osmolarity values of 16 eyes (group 1) who wore first generation SiH contact lenses were compared with those of 18 eyes (group 2) who wore second generation SiH contact lenses after three months follow-up. RESULTS: Before contact lens wear, tear film osmolarity of groups 1 and 2 were 305.02±49.08 milliosmole (mOsm) and 284.66±30.18mOsm, respectively. After three months of contact lens wear, osmolarity values were found 317.74±60.23mOsm in group 1 and 298.40±37.77mOsm in group 2. Although osmolarity values for both groups of SiH contact lens wear after three months periods were slightly higher than before the contact lens wear, the difference was not statistically significant. CONCLUSION: Contact lens wear may cause evaporation from the tear film and can increase tear film osmolarity leading to symptoms of dry eye disease. In the current study, there is a tendency to increase tear film osmolarity for both groups of SiH contact lens wear, but the difference is not statistically significant.

The relationship between the first episode of wheezing and matrix metalloproteinases-9 and MMP-2 and tissue inhibitors of MMP-1 levels in preterm infants.

AIMS: Matrix metalloproteinases (MMP) have been associated with neonatal lung morbidity and MMP dysregulation contributes to the pathology of chronic and acute lung disorders. Most of the previous studies were performed in the 1(st) weeks of life of the preterm newborns. There are no data on the serum levels of MMP-2, MMP-9 or tissue inhibitors of matrix metalloproteinases (TIMP-1) from preterm infants recovering from lung morbidities. We aimed to compare MMP-2, MMP-9 and TIMP-1 levels in preterm and term infants hospitalized with their first episode of wheezing. METHODS: We prospectively evaluated 18 preterm infants with a history of chronic lung disease, respiratory distress syndrome or oxygen therapy and 14 age- and sex-matched term infants who were admitted for a first episode of wheezing. We quantified total serum concentrations of MMP-2, MMP-9 and TIMP-1 to assess whether these serum markers levels were associated with the first episode of wheezing in infants with a history of oxygen therapy during the neonatal period. RESULTS: Upon hospitalization, MMP-2 and TIMP-1 levels were higher in preterm infants than in term infants. In contrast, there was no significant relationship between MMP-9 levels or the MMP-9/TIMP-1 ratio between preterm and term infants. The area under the receiver operating characteristic curve for MMP-2 was 0.70 (95% confidence interval [CI] 0.51-0.89). The area under the curve for TIMP-1 was 0.78 (95% CI 0.61-0.94). MMP-9, MMP-2 and TIMP-1 levels did not correlate with gestational age, gender or severity of wheezing. CONCLUSION: The negative proportion of MMP-9 to TIMP-1 that we detected in term infants was not present in preterm infants. The balance of MMP-9 to TIMP-1 may have been disrupted by lung damage in the premature infants. Overproduction of MMP-2 and TIMP-1 in the serum may be associated with the pathogenesis of wheezing in preterm infants.

Effects of levobupivacaine on wound healing.

BACKGROUND: Local anesthetic infiltration along the incision may be used to provide surgical anesthesia or postoperative analgesia. However, the effect of local anesthetics on wound healing remains controversial. In this investigation, we evaluated the effects of levobupivacaine on wound healing. METHODS: Sixty Wistar albino female rats weighing 230±20 g were included, with 10 rats in each group: group early c (early control): 3 mL isotonic saline; group early l1.25 (early levobupivacaine 1.25): 1.25 mg/kg per 3 mL levobupivacaine; group early l2.5 (early levobupivacaine 2.5): 2.5 mg/kg per 3 mL levobupivacaine; group late c (late control): 3 mL isotonic saline; group late l1.25 (late levobupivacaine 1.25): 1.25 mg/kg per 3 mL levobupivacaine; and group late l2.5 (late levobupivacaine 2.5): 2.5 mg/kg per 3 mL levobupivacaine. Rats in groups early c to early l2.5 were euthanized on the 8th day. Rats in groups late c to late l2.5 were euthanized on the 21st day. Wound tension strength, tissue hydroxyproline, and fibrotic index levels of the tissue samples from the early c and early l2.5 and late c and late l2.5 groups, respectively, on the 8th and 21st days were examined. RESULTS: Levobupivacaine decreased wound tension strength on the 8th day, especially a 2.5 mg/kg dose (P<0.001), and increased it on the 21st day (P<0.001). It also increased the inflammatory response (P<0.001) and collagen synthesis (8th day, P=0.109; 21st day, P=0.103) on both the 8th and 21st days. CONCLUSIONS: While levobupivacaine had a positive effect on wound healing during the early period, negative effects were observed thereafter. Additional studies at the molecular level are necessary to determine the cause of these apparently opposite effects.